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Applied and Environmental Microbiology, October 2008, p. 6333-6337, Vol. 74, No. 20
0099-2240/08/$08.00+0     doi:10.1128/AEM.02846-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Identification of Lacto-N-Biose I Phosphorylase from Vibrio vulnificus CMCP6{triangledown}

Masahiro Nakajima and Motomitsu Kitaoka*

National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan

Received 17 December 2007/ Accepted 13 August 2008

A β-1,3-galactosyl-N-acetylhexosamine phosphorylase (GalGlyNAcP) homolog gene was cloned from Vibrio vulnificus CMCP6. In synthetic reactions, the recombinant enzyme acted only with GlcNAc and GalNAc as acceptors in the presence of {alpha}-D-galactose-1-phosphate as a donor to form lacto-N-biose I (LNB) (Galβ1 -> 3GlcNAc) and galacto-N-biose (GNB) (Galβ1 -> 3GalNAc), respectively. GlcNAc was a much better acceptor than GalNAc. The enzyme also phosphorolysed LNB faster than it phosphorolysed GNB, and the kcat/Km for LNB was approximately 60 times higher than the kcat/Km for GNB. This result indicated that the enzyme was remarkably different from GalGlyNAcP from Bifidobacterium longum, which has similar activities with LNB and GNB, and GalGlyNAcP from Clostridium perfringens, which is a GNB-specific enzyme. The enzyme is the first LNB-specific enzyme that has been found and was designated lacto-N-biose I phosphorylase. The discovery of an LNB-specific GalGlyNAcP resulted in recategorization of bifidobacterial GalGlyNAcPs as galacto-N-biose/lacto-N-biose I phosphorylases.

* Corresponding author. Mailing address: National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan. Phone: 81-29-838-8071. Fax: 81-29-838-7321. E-mail: mkitaoka{at}affrc.go.jp

{triangledown} Published ahead of print on 22 August 2008.

Applied and Environmental Microbiology, October 2008, p. 6333-6337, Vol. 74, No. 20
0099-2240/08/$08.00+0     doi:10.1128/AEM.02846-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Nakajima, M., Nishimoto, M., Kitaoka, M. (2009). Characterization of Three {beta}-Galactoside Phosphorylases from Clostridium phytofermentans: DISCOVERY OF D-GALACTOSYL-{beta}1->4-L-RHAMNOSE PHOSPHORYLASE. J. Biol. Chem. 284: 19220-19227 [Abstract] [Full Text]  
  • Hidaka, M., Nishimoto, M., Kitaoka, M., Wakagi, T., Shoun, H., Fushinobu, S. (2009). The Crystal Structure of Galacto-N-biose/Lacto-N-biose I Phosphorylase: A LARGE DEFORMATION OF A TIM BARREL SCAFFOLD. J. Biol. Chem. 284: 7273-7283 [Abstract] [Full Text]  


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